Cell-cell signaling is an important component of the stem cell microenvironment, affecting both differentiation and self-renewal. However, traditional cell-culture techniques do not provide precise control over cell-cell interactions, while existing cell patterning technologies are limited when used with proliferating or motile cells. To address these limitations, we created the Bio Flip Chip (BFC), a microfabricated polymer chip containing thousands of microwells, each sized to trap down to a single stem cell. We have demonstrated the functionality of the BFC by patterning a 50×50 grid of murine embryonic stem cells (mESCs), with patterning efficiencies > 75%, onto a variety of substrates -a cell-culture dish patterned with gelatin, a 3-D substrate, and even another layer of cells. We also used the BFC to pattern small groups of cells, with and without cell-cell contact, allowing incremental and independent control of contact-mediated signaling. We present quantitative evidence that cell-cell contact plays an important role in depressing mESC colony formation, and show that E-cadherin is involved in this negative regulatory pathway. Thus, by allowing exquisite control of the cellular microenvironment, we provide a technology that enables new applications in tissue engineering and regenerative medicine.

In migrating cells, with especial prominence in lamellipodial protrusions at the cell front, highly dynamic connections are formed between the actin cytoskeleton and the extracellular matrix through linkages of integrin adhesion receptors to actin filaments via complexes of cytosolic "connector" proteins. Myosin-mediated contractile forces strongly influence the dynamic behavior of these adhesion complexes, apparently in two counter-acting ways: negatively as the cell-generated forces enhance complex dissociation, and at the same time positively as force-induced signaling can lead to strengthening of the linkage complexes. The net balance arising from this dynamic interplay is challenging to ascertain a priori, rendering experimental studies difficult to interpret and molecular manipulations of cell and/or environment difficult to predict. We have constructed a kinetics-based model governing the dynamic behavior of this system. We obtained ranges of parameter value sets yielding behavior consistent with that observed experimentally for 3T3 cells and for CHO cells, respectively. Model simulations are able to produce results for the effects of paxillin mutations on the turnover rate of actin/integrin linkages in CHO cells, which are consistent with recent literature reports. Overall, although this current model is quite simple it provides a useful foundation for more detailed models extending upon it.

Background: There have been many assumptions made about neuronal loss in mammals due to aging. However, when we examined the retinal ganglion cell layer of a marsupial, the quokka, from 0.5 to 13.5 years of age, we found that the total neuron number did not decrease significantly even into extreme old age. The retinal area increased slowly throughout life, leading to a decrease in cell density. Neuronal death in the rat retina has been assumed, since the cell density has been seen to fall with age. However, a similar study to ours in the quokka has never been performed in the laboratory rat, the model for so many experimental investigations. Objective: We decided to test the hypothesis that rats do not lose neurons due to aging and that an increase in retinal area, and not cell death, as previously suggested, might underlie the decreasing cell density seen in aging animals such as rats. Methods: We kept laboratory rats under standard, unvarying conditions throughout the trial, sacrificing 3 animals every 3 months up to 30 months, and examined the retinal ganglion cell layer through adult life. Results: We found that the numbers of neurons did not decrease in this species, even in the oldest rats. We also saw that the retinal area increased slowly with a concomitant slow decrease in mean neuronal density. Soma diameters of neurons gradually increased throughout life. Conclusions: When rats are kept under standard conditions, there is no neuronal loss in the retina during aging, although the cell density does decrease as a result of retinal expansion. It is not sufficient to measure retinal cell density to determine cell loss. In addition, it is important to know that in normal conditions there is no cell loss in the retinal ganglion cell layer as a result of aging. Any loss that is seen must be a result of additional factors.

A general statistical theory is developed for the masking effect of reverberation on the intelligibility of words. Speech is considered a series of discrete pulses distributed statistically over a 30-decibel range in sound pressure level in a given frequency band. The articulation index is calculated as a function of reverberation time, using preliminary values of speech pulse lengths and spacings obtained from Visible Speech spectrograms. The percent articulation for words is then calculated from the articulation index and is compared with Knudsen's experimental values. The theoretical values agree precisely with the measured values at reverberation times less than two seconds and differ by less than 17 percent out to six seconds. The calculations are extended to include a combination of background noise and reverberation.